【目的】研究NT3对早期卵泡存活、发育、激素合成的影响及其可能的作用机制,同时探讨NT3和LIF在早期卵泡启动发育方面是否存在协同或叠加效应。【方法】分离新生4d大鼠卵巢随机分配至新鲜组、基础培养组、NT3组、LIF组和NT3+LIF组,在37...【目的】研究NT3对早期卵泡存活、发育、激素合成的影响及其可能的作用机制,同时探讨NT3和LIF在早期卵泡启动发育方面是否存在协同或叠加效应。【方法】分离新生4d大鼠卵巢随机分配至新鲜组、基础培养组、NT3组、LIF组和NT3+LIF组,在37℃培养箱中培养14 d,经固定、包埋、切片后进行组织形态学、PCNA免疫组化、TUNEL凋亡分析,RTPCR检测KL m RNA表达及检测培养液中激素水平。【结果】相比新鲜组和基础培养组,NT3组、LIF组、NT3+LIF组中的始基卵泡比例下降,生长卵泡比例升高,同时PCNA蛋白表达显著增加;TUNEL分析显示NT3组、LIF组、NT3+LIF组的凋亡指数均显著降低;NT3、LIF组、NT3+LIF组卵巢中KL m RNA的表达量分别是对照组的(3.38±0.43)倍、(2.02±0.20)倍、(2.07±0.21)倍。【结论】NT3在早期卵泡存活、发育中有促进作用,上调KL基因表达可能是NT3的作用机制之一。NT3和LIF单独或相加均能促进卵泡发育,但两者之间无明显协同或叠加效应。展开更多
In the present study, we have cloned the gene of human neurotrophin3 (hNT3) from the genomic DNA of white blood cells (WBC) by polymerase chain reaction (PCR). The amplification products were cloned into pUC19 and seq...In the present study, we have cloned the gene of human neurotrophin3 (hNT3) from the genomic DNA of white blood cells (WBC) by polymerase chain reaction (PCR). The amplification products were cloned into pUC19 and sequenced. Genomic sequence comparison of the cloned fragment and the reported hNT3 (GenBank M61180) reveals 7 base differences: 1 in the signal peptide, 3 in the prepro peptide, and 3 in the mature hNT3. Except the 2 varied bases (16th, T to G; 285th, A to C) in the signal peptide and prosequence resulted in the change of their encoded aminoacids (TyrAsp; GlnHis), the other varied bases have no influence on their respective encoded aminoacids, and all the changes have no influence on the open reading frame (ORF) of the hNT3.展开更多
Objective Neurotrophin 3(NTF3)is involved in numerous biological processes;however,its role in hepatocellular carcinoma(HCC)is not well studied.This study investigated NTF3 function in HCC progression and revealed its...Objective Neurotrophin 3(NTF3)is involved in numerous biological processes;however,its role in hepatocellular carcinoma(HCC)is not well studied.This study investigated NTF3 function in HCC progression and revealed its underlying molecular mechanisms.Methods The prognostic relevance of NTF3 was determined through a bioinformatical analysis of publicly available TCGA data.Immunohistochemistry of HCC biopsies was performed to explore the expression of NTF3.Cell growth and proliferation were analyzed using a Cell Counting Kit-8(CCK-8)assay.Cell invasion and migration were analyzed using Boyden Transwell and wound healing assays.Protein expression and mRNA levels were evaluated through immunoblotting and quantitative polymerase chain reaction(qPCR).Cell apoptosis was evaluated with flow cytometry.Results NTF3 expression was significantly lower in HCC tissues than in adjacent non-tumor tissues.Low NTF3 expression was significantly associated with decreased patient survival and specific clinicopathological features.NTF3 overexpression reduced the proliferation,migration,and invasion abilities of HCC cell lines.Conclusion Decreased expression of NTF3 is associated with poor prognosis in HCC patients,likely due to its action in promoting HCC cell proliferation,migration,and invasion.Our findings provide a novel understanding into the pathogenesis of HCC and the role of NTF3 in tumor progression,suggesting that targeting NTF3 has potential therapeutic and diagnostic value for HCC.展开更多
Objective： To explore a new method to treat brachial plexus root avulsion experimentally by reimplantation combined with transplantation of neural stem cells （NSCs） modified by neurotrophin-3 gene （NT-3）. Method...Objective： To explore a new method to treat brachial plexus root avulsion experimentally by reimplantation combined with transplantation of neural stem cells （NSCs） modified by neurotrophin-3 gene （NT-3）. Methods： The total RNA was extracted from neonatal rat striatum and the NT-3 cDNA was obtained by reverse transcription and amplified by polymerase chain reaction. The NT-3 gene was transferred into NSCs via the pLEGFP-C 1, an expression plasmid vectors. The untransfected NSCs, the pLEGFP-C 1 treated NSCs, and the pLEGFP-C 1-NT-3 treated NSCs were transplanted into corresponding spinal cord segment with brachial plexus root avulsion. The survival, differentiation, and migration of the transplanted cells were determined under confocal laser scanning microscope or by immunohistochemistry method. The nerve regeneration was evaluated by gross observation, electrophysiological examination and reverse horseradish peroxidase tracing. Results： The NT-3 gene was successfully amplified and transferred into neural stem cells via the plasmid vectors. The transplanted cells survived, differentiated, and migrated and NT-3 was expressed within the spinal cord. The animals regained some muscle strength which was less than 3-degree muscular strength according to the British Medical Research Council （BMRC） evaluating system. The resuits of electrophysiological examination and reverse horseradish peroxidase tracing were superior in the pLEGFP-C 1- NT-3 group to the NSCs untransfected group or the pLEGFP- C1 group. Conclusion： Transplantation of NSCs modified by NT-3 gene combined with reimplantation is a relatively effective way to treat brachial plexus root avulsion experimentally. It still need further study to improve the results.展开更多
文摘【目的】研究NT3对早期卵泡存活、发育、激素合成的影响及其可能的作用机制,同时探讨NT3和LIF在早期卵泡启动发育方面是否存在协同或叠加效应。【方法】分离新生4d大鼠卵巢随机分配至新鲜组、基础培养组、NT3组、LIF组和NT3+LIF组,在37℃培养箱中培养14 d,经固定、包埋、切片后进行组织形态学、PCNA免疫组化、TUNEL凋亡分析,RTPCR检测KL m RNA表达及检测培养液中激素水平。【结果】相比新鲜组和基础培养组,NT3组、LIF组、NT3+LIF组中的始基卵泡比例下降,生长卵泡比例升高,同时PCNA蛋白表达显著增加;TUNEL分析显示NT3组、LIF组、NT3+LIF组的凋亡指数均显著降低;NT3、LIF组、NT3+LIF组卵巢中KL m RNA的表达量分别是对照组的(3.38±0.43)倍、(2.02±0.20)倍、(2.07±0.21)倍。【结论】NT3在早期卵泡存活、发育中有促进作用,上调KL基因表达可能是NT3的作用机制之一。NT3和LIF单独或相加均能促进卵泡发育,但两者之间无明显协同或叠加效应。
文摘In the present study, we have cloned the gene of human neurotrophin3 (hNT3) from the genomic DNA of white blood cells (WBC) by polymerase chain reaction (PCR). The amplification products were cloned into pUC19 and sequenced. Genomic sequence comparison of the cloned fragment and the reported hNT3 (GenBank M61180) reveals 7 base differences: 1 in the signal peptide, 3 in the prepro peptide, and 3 in the mature hNT3. Except the 2 varied bases (16th, T to G; 285th, A to C) in the signal peptide and prosequence resulted in the change of their encoded aminoacids (TyrAsp; GlnHis), the other varied bases have no influence on their respective encoded aminoacids, and all the changes have no influence on the open reading frame (ORF) of the hNT3.
基金Supported by grants from the National Natural Science Foundation of China(No.ZR2015HM025)Technology Development Program of Shandong Province,China(No.2011YD18003)CSCO-Hengrui Cancer Research Fund(No.Y-HR2018-293 and Y-HR2018-294).
文摘Objective Neurotrophin 3(NTF3)is involved in numerous biological processes;however,its role in hepatocellular carcinoma(HCC)is not well studied.This study investigated NTF3 function in HCC progression and revealed its underlying molecular mechanisms.Methods The prognostic relevance of NTF3 was determined through a bioinformatical analysis of publicly available TCGA data.Immunohistochemistry of HCC biopsies was performed to explore the expression of NTF3.Cell growth and proliferation were analyzed using a Cell Counting Kit-8(CCK-8)assay.Cell invasion and migration were analyzed using Boyden Transwell and wound healing assays.Protein expression and mRNA levels were evaluated through immunoblotting and quantitative polymerase chain reaction(qPCR).Cell apoptosis was evaluated with flow cytometry.Results NTF3 expression was significantly lower in HCC tissues than in adjacent non-tumor tissues.Low NTF3 expression was significantly associated with decreased patient survival and specific clinicopathological features.NTF3 overexpression reduced the proliferation,migration,and invasion abilities of HCC cell lines.Conclusion Decreased expression of NTF3 is associated with poor prognosis in HCC patients,likely due to its action in promoting HCC cell proliferation,migration,and invasion.Our findings provide a novel understanding into the pathogenesis of HCC and the role of NTF3 in tumor progression,suggesting that targeting NTF3 has potential therapeutic and diagnostic value for HCC.
基金This work was supported by grants from the National Natural Science Foundation of China （Grant No. 30471742）, Social Developmental Foundation of Jilin Province （Grant No.2001020）, Special Foundation of Ministry of Education （Grant No.20010183002）.
文摘Objective： To explore a new method to treat brachial plexus root avulsion experimentally by reimplantation combined with transplantation of neural stem cells （NSCs） modified by neurotrophin-3 gene （NT-3）. Methods： The total RNA was extracted from neonatal rat striatum and the NT-3 cDNA was obtained by reverse transcription and amplified by polymerase chain reaction. The NT-3 gene was transferred into NSCs via the pLEGFP-C 1, an expression plasmid vectors. The untransfected NSCs, the pLEGFP-C 1 treated NSCs, and the pLEGFP-C 1-NT-3 treated NSCs were transplanted into corresponding spinal cord segment with brachial plexus root avulsion. The survival, differentiation, and migration of the transplanted cells were determined under confocal laser scanning microscope or by immunohistochemistry method. The nerve regeneration was evaluated by gross observation, electrophysiological examination and reverse horseradish peroxidase tracing. Results： The NT-3 gene was successfully amplified and transferred into neural stem cells via the plasmid vectors. The transplanted cells survived, differentiated, and migrated and NT-3 was expressed within the spinal cord. The animals regained some muscle strength which was less than 3-degree muscular strength according to the British Medical Research Council （BMRC） evaluating system. The resuits of electrophysiological examination and reverse horseradish peroxidase tracing were superior in the pLEGFP-C 1- NT-3 group to the NSCs untransfected group or the pLEGFP- C1 group. Conclusion： Transplantation of NSCs modified by NT-3 gene combined with reimplantation is a relatively effective way to treat brachial plexus root avulsion experimentally. It still need further study to improve the results.
